Difference between revisions of "Part:BBa K404104"
| Line 24: | Line 24: | ||
<html> | <html> | ||
<p style="text-align:justify;"> | <p style="text-align:justify;"> | ||
| + | The BioBrick provided by the iGEM team Freiburg_Bioware 2010 can be used for producing recombinant viral particles by fusing the p5 TATA-less promoter downstream of the rep and cap open reading frames.<br /> | ||
In contrast to the natural location of the p5 promoter, the iGEM team Freiburg 2010 provides the RepCap plasmid with a relocated p5 promoter downstream of the RepCap genes (Figure 1). Additionally the p5 promoter lacks the TATA box element (AVIGEN 1997). Those modifications result in an attenuated expression of the larger Rep proteins therefore leading to normal transcription of the Rep proteins driven by p19 promoter and enhanced expression of the Cap proteins, which are under the control of the p40 promoter. Additionally, removing the p5 promoter downstream of the RepCap genes and deletion of the TATA box eliminates contamination with wtAAVs. Hence, alteration of the p5 promoter is useful for enhanced production of recombinant viral particles attenuating repression of Rep78/68 and improving gene transcription of the capsid proteins and Rep proteins involved in genome packaging. </p> | In contrast to the natural location of the p5 promoter, the iGEM team Freiburg 2010 provides the RepCap plasmid with a relocated p5 promoter downstream of the RepCap genes (Figure 1). Additionally the p5 promoter lacks the TATA box element (AVIGEN 1997). Those modifications result in an attenuated expression of the larger Rep proteins therefore leading to normal transcription of the Rep proteins driven by p19 promoter and enhanced expression of the Cap proteins, which are under the control of the p40 promoter. Additionally, removing the p5 promoter downstream of the RepCap genes and deletion of the TATA box eliminates contamination with wtAAVs. Hence, alteration of the p5 promoter is useful for enhanced production of recombinant viral particles attenuating repression of Rep78/68 and improving gene transcription of the capsid proteins and Rep proteins involved in genome packaging. </p> | ||
Revision as of 03:09, 25 October 2010
p5-TATAless promoter
| p5-TATA-less promoter | |
|---|---|
| |
| BioBrick Nr. | K404104 BBa K404104 |
| RFC standard | RFC 10 |
| Requirement | pSB1C3 |
| Source | pAAV_RC from Stratagene |
| Submitted by | [http://2010.igem.org/Team:Freiburg_Bioware FreiGEM 2010] |
The BioBrick provided by the iGEM team Freiburg_Bioware 2010 can be used for producing recombinant viral particles by fusing the p5 TATA-less promoter downstream of the rep and cap open reading frames.
In contrast to the natural location of the p5 promoter, the iGEM team Freiburg 2010 provides the RepCap plasmid with a relocated p5 promoter downstream of the RepCap genes (Figure 1). Additionally the p5 promoter lacks the TATA box element (AVIGEN 1997). Those modifications result in an attenuated expression of the larger Rep proteins therefore leading to normal transcription of the Rep proteins driven by p19 promoter and enhanced expression of the Cap proteins, which are under the control of the p40 promoter. Additionally, removing the p5 promoter downstream of the RepCap genes and deletion of the TATA box eliminates contamination with wtAAVs. Hence, alteration of the p5 promoter is useful for enhanced production of recombinant viral particles attenuating repression of Rep78/68 and improving gene transcription of the capsid proteins and Rep proteins involved in genome packaging.
Figure 1: T: p5 TATA-less promoter is located downstream of the rep and cap ORF..
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 98
Illegal BsaI site found at 135